MPB 64 Possesses Tuberculosis‐Complex'‐Specific B‐ and T‐Cell Epitopes

Abstract

We have developed monoclonal antibodies (MoAb) reactive with a protein from Mycobacterium tuberculosis of apparent molecular mass 24 kDa. This protein was shown to be identical with MPB 64 (Harboe et al.,) MoAb bound to four different epitopes of which two were restricted to the 'tuberculosis complex' and two were also found in mycobacteria not belonging to the 'tuberculosis complex'. The cross-reactive MoAb demonstrate that MPB 64 is present in more mycobacterial species than previously assumed. MPB 64 was shown to induce strong delayed type hypersensitivity (Dth) reactions in outbred guinea pigs immunized with M. tuberculosis and M. bovis bacille Calmette-Guérin (BCG). No reaction was observed in animals immunized with mycobacteria not belonging to the 'tuberculosis complex'. The Dth-inducing capacity of MPB64 was compared with that of another 24 kDa protein purified from M. tuberculosis and of the previously described 38 kDa protein. The Dth responses to these three antigens were further analysed in four inbred guinea pig strains. A genetic restriction of the ability of the animals to respond to MPB 64 as well as to the 38 kDa protein was observed.

Modulation of the Bovine Delayed-Type Hypersensitivity Responses to Defined Mycobacterial Antigens by a Synthetic Bacterial Lipopeptide

“…However, investigation of such antigens as bovine skin test reagents has not been reported previously and therefore was the subject of the present study. ESAT-6, MPB64, and MPB70 have previously been shown to be capable of stimulating DTH responses in guinea pigs that were infected or sensitized with either live Mycobacterium tuberculosis, M. bovis BCG, or heat-killed M. bovis strains (1,11,15,18,31,34). The demonstration that recombinant forms of these antigens also induced DTH in M. bovis-infected guinea pigs in the present study confirmed that DTH-inducing antigens may be useful as skin test reagents in cattle.…”

Molecular composition of the outermost capsular material of the tubercle bacillus

“…Seven-day short-term culture filtrate of M. tuberculosis was produced essentially as described by P. Andersen et al (1991). After day 7, the bacterial cells were removed from the growth medium by filtration (Milliguard Housing system, Millipore) and the culture filtrate was concentrated (~1 0 0 ) on an Amicon YM3 membrane resulting in protein concentrations ranging from 5 to 10 mgml-'.…”

“…B. Andersen et al, 1991); the mAbs were diluted in PBS (pH 7.6) containing 0.37 M NaCl and 0-05 YO (w/v) Tween 20 (culture supernatants were diluted 1 :25, and protein A culture supernatant mAbs were diluted 1 : 500). Bound mAbs were detected by using horseradish-peroxidase-coupled rabbit anti-mouse immunoglobulins (P216; Dakopatts) at a dilution of 1 : 1000.…”

Identification and Characterization of the Protective Gene of homolog of Exported Protein 1

“…Substantial evidence indicates that exported molecules can affect virulence and influence immunity to infection. They may act as virulence factors, as illustrated for Yersinia enterocolitica (39), Bordetella pertussis (40), and Strongyloides stercoralis (41), or as major targets of protective immunity, as illustrated for Mycobacterium tuberculosis (42)(43)(44). This apparent high immunogenicity of exported proteins may perhaps be attributable to the increased accessibility to proteolysis and transportation associated with targeting to the cytoplasm and more efficient entry into the class I and class II pathways (34 -37).…”